Identification and Differentiation of Poultry Meat and Products Using PCR-RFLP Technique

Abstract

The mitochondrial cyt b gene plays a serious role in investigating untruthful meat species. This study aimed to authenticate the species of poultry products (Escallop, Nugget, Steak, and Sausage) depending on cyt b gene by using universal cyt b primer. DNA was isolated, and then a band of 359 bp of a mitochondrial cytochrome b gene was produced during the PCR amplification. The PCR products were exposed to Hinf1 and Rsa I restriction enzymes. The restricted fragments produced by restriction fragment length polymorphism technique (RFLP), were run by agarose gel electrophoresis. Results showed that all products had a similar band except sausage product does not follow the rule and showed mislabeling product by the REs, Two bands were yielded by HinfI RE for all products (114 and 245) bp with the differentiated sausage among other products based on the fake product (63 and 296) bp, while digestion by Rsa I produced three bands for escallop, nugget, and steak, (63, 100, 196), but only two bands for sausage was generated (148 and 211). As result, the study offered that analyzing meat products to detect the origin species via a PCR-RFLP technique by using these restriction enzymes can give reliable results. In short sausage is considered as fraud products because the results showed different bands as compared with poultry meat.

Molecular Detection of Raw Meat for Some Animal Species Using RFLP-PCR Technique

“…In this study mt cyt b gene 359bp was used to identification and investigation of origin species of animal meat such as (sheep, goat, cattle and donkey) since this short mitochondrial fragment is repeated the widest range in nucleotide databases (Guo et al, 2005). Amplification of the pair primers of mt cyt b gene by PCR produced 359bp in all extracted DNA animal samples this confirms the more benefits of using high copy number mtDNA over nuclear DNA, which is approved by previous study (Khan et al, 2018;Dilger et al, 2020 andTaha et al, 2021).…”

“…The study for detection of species origin depended on the mt cyt b gene have ranged between less than 400bp to more than 900bp RFLP-PCR DNA sequencing and variable size species specific multiplex PCR (Lee et al, 2009). A replacement DNA identification method is depended on the PCR amplification of a piece of the mt cyt b gene then digestion the amplicons by restriction enzymes via RFLP analysis, which produce species specific pattern, this technique requires less equipment and cost than sequencing (Ahmed, 2007;Abdel-Rahman et al, 2009 andTaha et al, 2021). Therefore, the objective of this study was to use RFLP-PCR analysis of the mt cyt b gene, using 2 restriction enzymes (HinfI and Rsa I) for differentiation of sheep, goat, cattle and donkey species in Erbil Kurdistan region.…”

Molecular Identification of the American Cockroach (Periplaneta americana) in the Kurdistan Region of Iraq (Dictyoptera: Blattidae)

“…According to [29][30][31], molecular research identifies new and cryptic species, which are difficult to identify depending only on morphological features. Morphological identification performed insufficiently compared to molecular markers [32]. Proper identification is the initial stage in pest management, and DNA-based techniques are the most effective approach for identifying insect species from every position.…”

Comparison of different DNA preparatory methods for development of a universal direct PCR-RFLP workflow for identification of meat origin in food products

“…Within the mitochondrial genes, 16S rRNA has been used for broad range of mammalian and birds' species because of its evolutionary stability (Vences et al, 2016;Ha et al, 2017;Lalitha & Chandavar, 2017). Furthermore, the developed direct PCR method is coupled with RFLP assay which is much more desirable for molecular based meat identification, especially, in cases where large number of samples have to be processed as it does not require DNA sequencing and/or specialized equipment and reduces the cost and time for post-PCR processing of samples (Guan et al, 2019;Gargouri et al, 2021;Vaithiyanathan et al, 2021;Taha et al, 2021). It has been mostly applied for species discrimination in processed and unprocessed meat products because of its simplicity, quicker detection and reduced cost (Al et al, 2020;Asghar et al, 2022;Farag et al, 2022).…”